Some metabolic differences between human skin and aponeurosis fibroblasts in culutre

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2-Deoxy-D-glucose (2-DOG) uptake was studied in skin fibroblast cultures from control children and children with Alagille syndrome or syndromic paucity of interlobular bile ducts (PILBD). No significant differences in uptake were observed between patients and controls. However, as the scatter of the results was larger in the fibroblasts from patients, we attempted to establish for these patients a relationship between 2-DOG uptake and some biochemical parameters. We observed an inverse relationship between this uptake and the levels of plasma cholesterol and phospholipids (r = -0.85). Compared to controls, 2-DOG uptake was significantly lower in cultures from patients who had very high levels of cholesterol (P2 group), but not in cultures from patients with moderately increased levels of cholesterol (P1 group). The level of total cellular cholesterol in cultured cells from the P1 and P2 groups was not significantly different from the control level, but we found marked differences between the concentrations of fatty acids. In the cultures from patients (especially the P2 group), we observed a significant increase in total fatty acids; among the saturated fatty acids, this increase chiefly concerned the 18:0 (14%) and among the polyunsaturated the n - 3 fatty acids (55%). The high concentrations of 20:5, 22:5 and 22:6, which enhance membrane fluidity, might explain the decrease in 2-DOG uptake found in the cultures from patients (P2 group) with PILBD. The nature of these abnormalities might be connected with the genetic origin of Alagille syndrome.

Section: Cell Culturesmentioning

confidence: 99%

2‐Deoxy‐d‐glucose uptake and fatty acid content in fibroblast cultures from children with syndromic paucity of interlobular bile ducts (alagille syndrome)

Couturier

Lemonnier

1990

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“…Strains of diploid skin fibroblasts (SF) were initiated from explants as previously described (Lemonnier et al 1980) and subcultured at 37°C in 25 cm z plastic tissue culture flasks (T25, Falcon) with 4ml of medium and under 5% CO2 in air. Basic medium was MEM containing 5.5mmol/L glucose, supplemented with 2mmol/L glutamine, antibiotics and 10% fresh non-pooled human serum (HS).…”

Section: Methodsmentioning

confidence: 99%

Impaired hexose uptake by diploid skin fibroblasts from galactosaemic patients. Connection with cell growth and amino acid metabolism, and possible bearing on late‐onset clinical symptoms

Wolfrom

Raynaud

Kadhom

et al. 1992

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In skin fibroblasts of patients presenting with galactosaemia, either from galactose 1-phosphate uridyltransferase or galactokinase deficiency, a deficit in extracellular glucose utilization was observed. This deficit was constant over 3 weeks of continuous cell growth in a medium containing 5.5 mmol/L glucose as the only hexose, and homologous serum. Levels of glucose utilization by deficient skin fibroblasts were stable at about 65-70% of the glucose utilization of control normal skin fibroblasts. Cell morphology was normal, and cell growth was subnormal during this period. However, the energy provision appeared sufficient for cellular needs since cell growth in this glucose medium was observed not to depend on the presence of extracellular glutamine. In contrast, glutamine was required for growth of galactosaemic fibroblasts cultured in medium containing 5.5 mmol/L galactose. If expressed in many cell types, this impaired glucose uptake would be expected seriously to damage highly glucose-dependent tissues such as the central nervous system. This might be of relevance to the persistent neurological damage observed in many galactosaemic patients in spite of their compliance with an early strict galactose-free diet.

“…Skin fibroblasts were obtained from eleven healthy controls and three patients with prolidase deficiency (one 35-year-old man and two women aged 20 and 23 years respectively). The culture initiation and subcuttivation techniques were as described previously (Lemonnier et al, 1980). Cells were harvested between days 10 and 14 MS received 21.7.87 Accepted 6.4.88 by trypsinization and centrifuged at 400 g for 5 min.…”

Section: Methodsmentioning

confidence: 99%

Prolinase activity in prolidase‐deficient fibroblasts

Miech

Myara

Mangeot

et al. 1987

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The activity of prolinase (EC 3.4.13.8) was studied in cultured skin fibroblasts derived from three patients with deficient prolidase (EC 3.4.13.9). With pro-val as substrate and manganese in the reaction buffer, prolinase activity was higher in prolidase-deficient cells than in control cells (mean (SEM) 917 (67) nmol min-1 mg-1, n = 3, control mean (SEM) 294, (50), n = 11). The Michaelis constants were not different for the pro-val and progly substrates in control and prolidase deficient fibroblasts. However, the constants for Vmax rose for both substrates in deficient cells. These results demonstrate that prolinase activity increases in prolidase-deficient fibroblasts as also shown in the plasma of patients with prolidase deficiency. We suggest that in prolidase-deficient fibroblasts, this rise in prolinase activity constitutes an attempt to compensate for the prolidase deficiency by increasing the greatly reduced intracellular proline pool.